绿色荧光蛋白与黄瓜花叶病毒运动蛋白融合基因的构建及在大肠杆菌中的表达

摘要

A fusion gene of green fluorescent protein(GFP)and movement protein of cucumber mosaic virus(CMV MP)was obtained by using recombination-PCR.The procaryotic expression vector pKENG-M for GFP:CMV MP was constructed by cloning in pKEN.Results from SDS-PAGE and western blot analysis showed that the 57 kD protein encoded by the fusion gene was expressed correctly in E.coli DH5　.Green fluorescence in the bacteria was observed under a confocal microscope when excited with 488 nm laser line,indicating GFP activity was maintained.%采用重组-PCR方法，扩增获得绿色荧光蛋白（GFP）与黄瓜花叶病毒运动蛋白（CMV MP）融合基因，将其克隆到pKEN上，构建了该融合基因的原核表达载体pKENG-M。SDS-PAGE及免疫印迹分析显示，57 kD的融合蛋白基因在大肠杆菌DH5中正确表达。激光共聚焦显微镜分析表明，在488 nm光激发下，菌体呈亮绿色，表明融合蛋白保持绿色荧光特性。实验结果为进一步研究CMV MP的功能及其与胞间连丝和其他细胞成分的作用关系奠定了基础。